Process of inhibiting microorganisms with alkylene bis-dithiocarbamate esters



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Bit? 2 a 97a 9 082?. \a gaeas r i i Unite PROCESS OF INHIBITINGMICROORGANISMS glsgllkLKYLENE BIS-DITHIOCARBAMATE No Drawing. Filed Apr.25, 1 957, Set. No. 654,982

6 Claims. 01. 16722) This invention relates to fungical, bactericidaland germicidal organic compounds and more particularly to fungicidescomprising bis-dithiocarbamate esters and the methods of preparing thesame. Tins is a continuationin-part application of application SerialNo. 514,105, filed June 8, 1955, now abandoned.

It is an object of this invention to provide organic compoundscomprising alkylene bis-dithiocarbamate esters having new and morepotent fungicidal properties than known fungicides.

Another object of this invention is to provide organic compoundscomprising esters of alkylene bis-dithiocarbamic acid which havemicro-organism-destroying properties and which retain such propertiesover long periods of time.

Still another object of this invention is to provide a fungicide whichexhibits excellent activity against many fungi.

In attaining the objects of this invention, one feature resides inreacting a di-metallic salt of a bis-dithiocarbamate with an organichalide to form a bis-dithiocarbamate ester having these excellentfungicidal properties.

Another features resides in reacting a dimetallic salt of2-hydroxytrialkylene-1, S-bis-dithiocarbamate with an organic halide toform a 3-bis-dithiocarbamate ester having excellent micro-organismdestroying properties.

Other objects, features and advantages will become more apparent fromthe following description of the invention.

It has been found that the condensation reaction product formed byreacting a dimctallic salt of a bis-dithiocarbamate with an organichalide has the properties of easily and quickly destroying variousbacteria or fungi which are treated therewith.

In particular, it has been found that a compound having the formulawherein R is a radical selected from the group consisting of saturatedand unsaturated hydrocarbons, saturated and unsaturated substitutedhydrocarbons, and substituted and unsubstituted carbocyclic radicals, Ris a radical selected from the group consisting of alkylene, substitutedalkylene, primary amines and secondary amines, and R is a radicalselected from the group consisting of hydrogen and alkyl, exhibitsconsiderably stronger fungicidal and bactericidal activity thanundecylenic acid when tested against various microorganisms by the paperdisk technique.

Examples of radicals from which R may be selected 2,974,082 PatentedMar. 7, 1961 allyl, substituted allyl, aralkyl, substituted aralkyl,aryl, substituted aryl, phenacyl, acetyl, benzoyl, cyclohexyl,polyethylene glycol [H(OCH CH and others.

Examples of radicals from which R may be selected include, among others,ethylene, propylene, butylene, etc., Z-hydroxy propylene, diamine andother amines wherein the nitrogen atom has at least one availablehydrogen atom.

R may be H, or radicals such as methyl, ethyl, and the like.

A compound having the above formulation has also been found to have apotent germicidal effect upon Microsporum Ianosum, Staphylococcusaureus, Microsporum audouini, Microsporum gypseum, Achorian schoenleini(Trichophyion schoenleini), and T richophyton rubrum.

The new bis-dithiocarbamate esters which exhibit these potent fungicidaland bactericidal properties are obtained by the condensation reaction oftwo molecules of an organic halide with one molecule of a dimetallicbisdithiocarbamate. The condensation products thus formed are onlyslightly soluble in water at 25 C., but may be suspended in water by useof suspending agents such as the Tweens. TliFTweens are non-ionicsurface active agents whichare commercially available and are derived byadding polyoxyethylene chains to the nonesterified hydroxyls of thepartial esters of the common fatty acids and hexitol anhydrides derivedfrom sorbitol. These condensation products are also soluble in varyingdegrees in organic solvents such as the alcohols, ethers, acetone,benzene, etc.

The following examples illustrate several embodiments of the process forpreparing the esters of bis-dithiocarbamates.

EXAMPLE 1 Bis-Z-hydroxyethyl ethylene bis-dithiocarbamate To aconstantly stirred mixture prepared from 0.25 mole of disodium ethylenebis-dithiocarbamate and 200 ml. of water is added 0.5 mole of ethylenechlorohydrin slowly over a period of 20-30 minutes. After addition ofall the ethylene chlorohydrin the reaction mixture is stirred for 3.5hours keeping the temperature between 30-40 C. At this point thereaction mixture is extracted with several portions of ether. Thecombined ether extracts are dried over anhydrous sodium sulfate and theether distilled. The crude material remaining is triturated rapidly withpetroleum ether until crystallization takes place. The yellow coloredcrude product is purified by recrystallization from ethanol and water inthe form of fine yellow crystals having a melting point of 71-73 C.

EXAMPLE 2 Bis-3-hydroxypropyl ethylene bis-dithiocarbamate To aconstantly stirred mixture prepared from 0.25 mole of disodium ethylenebis-dithiocarbamate and 200 ml. of water is added 0.5 mole oftrimethylene chlorohydrin slowly over a period of 20-30 minutes. Afteraddition of all the trimethylene chlorohydrin the reaction mixture isstirred for 3.5 hours. The reaction mixture is then cooled to 0 C. andvery viscous oil separates from solution. The water is decanted oil andthe oil dissolved in warm isobutyl methyl carbinol and dried overanhydrous sodium sulfate and filtered. The isobutyl include certainlower alk ls substituted thyl carbinol is extracted with petroleum etherand the petroleum ether remaining in the ester layer is removed undervacuum. The remaining purified product is a viscous yellow oil.

EXAMPLE 3 Di-n-propyl ethylene bfs-dithiocarbamate To a constantlystirred mixture prepared from 0.25 mole of disodium ethylenebis-dithiocarbamate and 200 ml. of water is added 0.5 mole of n-propylbromide slowly over a period of 25-30 minutes. After addition of all then-propyl bromide the reaction mixture is stirred for 3.5 hours. Thereaction mixture is then cooled to C. and a yellow crystallineprecipitate separates. The precipitate is collected on a filter andpurified by recrystallization from ether in the form of bright yellowcrystals melting at 44-46 C.

EXAMPLE 4 Diallyl ethylene bis-dithiocarbamate To a constantly stirredmixture prepared from 0.25 mole of disodium ethylene bis-dithiocarbamateand 200 ml. of water is added 0.5 mole of allyl chloride, slowly over aperiod of 25-30 minutes keeping the mixture at 3040 C. After addition ofall the allyl chloride the reaction mixture is stirred for 1.5 hours.The reaction mixture is then extracted with several portions of ether.The combined ether extracts are dried over anhydrous sodium sulfate andfiltered. The ether is evaporated with the last small amounts beingremoved under vacuum. The remaining purified product is a bright yellowviscous oil with an odor similar to garlic.

EXAMPLE 5 Dibenzyl ethylene bis-dithiocarbamate To a constantly stirredmixture prepared from 0.25 mole of disodium ethylene bis-dithiocarbamateand 200 ml. of water is added 0.5 mole of benzyl chloride slowly over aperiod of 25-30 minutes keeping the mixture at 30-40" C. After theaddition of all the benzyl chloride the reaction mixture is stirred for2 hours and a light yellow precipitate separated. The precipitate isfiltered and washed with 150 ml. of water. The product is purified byre-crystallization from acetone and water from which is obtained lightyellow crystals of melting point 114-116 C.

EXAMPLE 6 Bis(acetylmethyl) ethylene bis-dithiocarbamate In a 500 ml.,three-necked flask, fitted with a mechanical stirrer, dropping funneland thermometer, was placed a mixture of 64 gm. (0.25 mole) of disodiumethylene bis-dithiocarbamate and 150 ml. of water. Chloro-2- propanone,46 gm., (0.50 mole) was added slowly over a period of ten to fifteenminutes to the constantly stirred mixture which was kept at atemperature of 30-40 C. After the addition of the chloro-2-propanone,the reaction mixture was stirred for twenty-five to thirty minutesduring which time a light-tan precipitate separated. The precipitate wascollected by filtration and washed twice with 50 ml. portions of coldwater. The product was purified by re-crystallizing three times from acyclohexanone n-pentane mixture. The product was then dried in vacuumfor three hours at 60 C. The yield was 22 gm. (54.3%) of the ester whichmelted at 230- 232 C. with decomposition.

Analysis.-Calculated N 8.64%.

EXAMPLE 7 Bis(f0rmylmethyl) ethylene bis-dithiocarbamate In a 500 ml.,three-necked flask fitted With a mechani- Found N 8.55%.

cal stirrer, dropping funnel and thermometer was placed a mixture of 64gm. (0.25 mole) of disodium ethylene bis-dithiocarbamate and 150 ml. ofwater. Chloro-acetaldehyde, 39.25 gm. (0.50 mole), was added slowly overa one hour period to the constantly stirred mixture which was maintainedat a temperature of 510 C. At the end of this time a tan syrupy mass hadseparated along with a smaller amount of a brown tarlike ma terial. Thetan material was dissolved in acetone, de colorized with charcoal andthen re-crystallized three times from an acetone-water solution. Theproduct was then dried in vacuum at 40 C. for twenty-four hours. Theyield was 12 gm. (16.3%) of the ester which melted at 170 C. withdecomposition.

Analysis-Calculated N 9.46%.

EXAMPLE 8 Bis(carbeth0xymethyl) ethylene bis-dithiocarbamate In a 500ml., three-necked flask, fitted with a me- Found N 9.53

' chanical stirrer, dropping funnel and thermometer, was

placed a mixture of 64 gm. (0.25 mole) of disodium ethylenebis-dithiocarbamate and 150 ml. of water. Ethyl bromoacetate, 83.5 gm.(0.50 mole), was added slowly over a thirty-minute period to theconstantly stirred mixture which was maintained at a temperature of 10-15 C. Stirring was continued for one hour, during which time thereaction mixture became neutral to litmus and a light-yellow precipitateseparated. The precipitate was collected by filtration and washed twicewith 50 ml. of cold water. The product was re-crystallized three timesfrom an acetone-n-pentane solution. The ester was a fluffy, white,crystalline material. The yield was 30 gm. (31.3%) of the ester whichmelted at 121 123 C.

Analysis.--Calculated N 7.28%.

EXAMPLE 9 Bis(p0lyethylene glycol) ethylene bis-dithiocarbamate (aver.M.W. 560) In a 500 ml., three-necked flask, fitted with a mechanicalstirrer, dropping funnel and thermometer, was placed a mixture of 64 gm.(0.25 mole) of disodium ethylene bis-dithiocarbarnate and 150 ml. ofwater. Polyethylene glycol chloride 210*, gm. (0.50 mole), was added inone lot. The reaction mixture was then heated to 65 C. and stirredconstantly for three hours after which it was then cooled to 50 C. Theester was then separated by salting out with sodium chloride. The esterwhich was a dark-brown oil, was then dissolved in cyclohexanone, driedover anhydrous sodium sulfate for forty-eight hours and filtered. Thecyclohexanone was then removed from the ester by repeated extractionwith n-pentane. After separation, the ester was dried in vacuum at 6065C. The yield was 60 gm. (42.8%) of the ester.

Analysis.--Calculated N 5.01%.

EXAMPLE 10 Bis(p0lyethylene glycol) ethylene bis-dithiocarbamate (aver.M. W. 960) In a 500 ml., three-necked flask, fitted with a mechanicalstirrer, dropping funnel and thermometer, was placed a mixture of 64 gm.(0.25 mole) of disodium ethylene bis-dithiocarbamate and 100 ml. ofwater. Polyethylene glycol chloride 410*, 205 gm. (0.50 mole), was addedin one lot. The reaction mixture was heated to 70 C. and stirredconstantly for four hours. Cyclohexanone, ml. was then added to thereaction mixture and the sodium Found N 7.26%.

Found N 4.78%

Product 0! Carbide and Carbon Chemicals Co.

chloride was removed by filtration. 'Ihe cyclohexa'noneester solutionwas then dried over anhydrous sodium sulfate for forty-eight hours andfiltered. The cyclohexanone was then removed from the ester by repeatedextraction with n-pentane. After separation the ester was dried invacuum at 65 7 0 C. for four hours. The yield was 203 gm. (84.5% )of theester.

Analysis-Calculated N 3.1%. Found N 3.0%.

EXAMPLE 11 Bis(2-methylallyl) ethylene bis-dithiocarbamate In a 500 ml.,three-necked flask, fitted with a mechanical stirrer, dropping funneland thermometer, was placed a mixture of 64 gm. (0.25 mole) of disodiumethylene bis-dithiocarbamate and 150 ml. of water. To this constantlystirred mixture was added 45.25 gm. (0.50 mole) of 3-chloro-2-methy1propene over a period of twenty minutes. The mixture was then heatedslowly to 45 C. and stirring continued for two hours, during which timea yellow metastable oil separated. The oil was removed and washed twicewith 50 m1. portions of water and then dried in vacuum at 40 C. forforty-eight hours, during which time crystallization took place. Theproduct was purified by recrystallizing twice from acetone. The yieldwas 27 gm. (33.8%) of the ester, a pale-yellow material melting at58-60.

Analysis.Calculated N 8.75%

EXAMPLE 12 Bis(p-chl0r0benzyl) ethylene bis-dithiocarbamate Found N8.76%

In a 500 ml., three-necked flask, fitted with a mechanical stirrer,dropping funnel and thermometer, was placed a mixture of 51.2 gm. (0.20mole) of disodium ethylene bis-dithiocarbamate and 150 m1. of water. Tothis mixture was added 64.4 gm. (0.40 mole) of p-chlorobenzyl chloride.The reaction mixture was heated to 60 C. and stirred constantly for twohours, during which time a precipitate separated. The precipitate wascollected by filtration, washed with three 50 ml. portions of water,then .rir dried. The product was then recrystallized four times from asolution of dioxane and n-pentane. The ester was a white, crystallinematerial. The yield was 24 gm. (24.2%) of the ester which melted at143-145 C.

Ar.talvsis.Calculated N 5.64%. Found N 5.64%.

EXAMPLE 13 Bis(2,4-dichl0r0benzyl) ethylene bis-dithiocarbamate In a 500ml., three-necked flask, fitted with a mechanical stin'er, droppingfunnel and thermometer, was placed a mixture of 51.2 gm. (0.20 mole) ofdisodium ethylene bisdithiocarbamate and 150 ml. of Water. To thismixture was added 78.2 gm. (0.40 mole) of 2,4-dichloro-benzyl chloride.The reaction mixture was heated to 60 C. and stirred constantly for fourhours during which time an oily material separated. The oil wascollected by decanting the mother liquid and washed three times withwater. After drying the oil in vacuum, it was triturated rapidly withn-pentane for two to three hours, during which time crystallization tookplace. The ester was then re-crystallized four times from a solution ofdioxane and n-pentane. The ester was a yellow, crystalline productmelting at 1101l2 C. The yield was 13 gm. (12.3%).

Analysis.Calculated N 5.30%. Found N 5.30%.

EXAMPLE l4 Bis(3,4-dichl0r0benzyl) ethylene bis-dithiocarbamate In a 500ml., three-necked flask fitted with a mechanical stirrer, droppingfunnel and thermometer, was placed a chloride. The reaction mixture washeated to 60 C. and stirred constantly for three hours, during whichtime a thick yellow oil separated and the mixture became neutral tolitmus. The oil was collected by decanting the mother liquor and thenwashed three times with 50 m1. portions of water. The product was driedin vacuum for fortyeight hours and then triturated rapidly for two tothree hours with n-pentane, during which time crystallization tookplace. The product was then re-crystallized four times from a solutionof dioxane and n-pentane. The ester was a yellow, crystalline productmelting at 118-120 C. The yield was 35 gm. (33%).

Analysis.--Calculated N 5.30%. Found N 5.29%.

EXAMPLE A Disodium-2-hydr0xytrimethylene-1, 3-bis-dz'thi0carbamate In a500 ml., three-necked flask, a solution of 45.06 gm. (0.50 mole) of1,3-diamino-2-propanol in ml. of ethanol was cooled in an ice-water bathto 30 C. A solution of 40 gm. (1.0 mole) of sodium hydroxide in 40 ml.of water was cooled to to 30 C. and added in one EXAMPLE 15Dibenzyl-Z-hydroxytrimethylene-I, 3-bis-dithiocarbamate In a 500 ml.,three-necked flask, fitted with a mechanical stirrer, dropping funneland thermometer, was placed a solution of approximately 0.25 mole ofdisodium-2-hydroxytrimethylene-l, 3-bis-dithiocarbamate in 150 ml. ofwater. To this was added 0.50 mole of benzyl chloride dropwise over afifteen to twenty minute period while stirring constantly. Stirring wascontinued and the temperature of the reaction mixture rose to 45 C.After one hour, a dark-green mass had precipitated from solution. Theprecipitate was separated by filtrationand washed twice with 150 ml.portions of water. The product was then dissolved in acetone andre-crystallized by addition of n-pentane. This was repeated three timesand the product was then dried in vacuum at 60 C. for four hours. Theester was a very viscous yellow oil with the odor of leek. The yield was57 gm. (57%).

Analysis.Calculated N 6.65%. Found N 6.70%.

EXAMPLE 16 Bis( acetylm ethyl -2-hydr0xytrimethylene-1,3-bis-dith-i0carbamate In a 500 ml., three-necked flask fitted with amechanical stirrer, drop-ping funnel and thermometer, was placed asolution of approximately 0.25 mole of disodium-2-hydroxytrimethylene-l, 3-bis-dithiocarbamate in 150 ml. of alcoholand water. To this constantly stirred solution, maintained at 25 C., 46gm. (0.5 mole) of chloro-2-pro panone was added dropwise over a thirtyminute period. The reaction was completed at this time and the producthad separated as a dark green metastable oil at the bottom of thereaction flask. The mixture was cooled at 10 C. and the mother liquorpoured olf. The oil was dissolved in acetone and water was then addeduntil the solution was slightly cloudy. After 36 hours the productseparated as a tan, crystalline solid. The ester was collected byfiltration, air dried and then re-crystallized three times from amixture of cyclohexanone and n-pentane. It was then dried in a vacuumoven for seventy-two hours 7 at 40 C. The yield was 49 gm. (55.4%). Themelting point was 151-153 C.

Analysis-Calculated N 7.91%.

EXAMPLE 17 Bis( carbethoxymethyl) -2-hydroxytrimethylene-I3-bis-dithiocarbamate Found N 8.33%.

. cooled to C. and the mother liquor poured off. The

product was washed twice with 50 ml. portions of water, air dried andre-crystallized three times from a mixture of cyclohexanone andn-pentane. It was then dried in vacuum for seventy-two hours at 40 C.The yield was gm. (19.3%) of the ester which melted at 151- 153 C.

Analysis.Calculated N 6.8%. Found N 7.3%.

EXAMPLE B Sodium dithiocarbanilate In a 500 ml., three-necked flask, asolution of 93 gm. (1.0 mole) of aniline in 150 ml. of ethanol, wascooled in an ice-water bath of C. A solution of 40 gm. (1.0 mole) ofsodium hydroxide in 40 ml. of water was cooled to C. and added in onelot to the cooled solution of aniline. The flask was then fitted with areflux condenser, mechanical stirrer and a dropping funnel. Cooling wascontinued and 80 gm. (1.05 moles) of carbon disulfide was added dropwiseover a period of one hour with constant stirring. After the addition ofthe carbon disulfide the reaction mixture was stirred vigorously forfour hours. At this time 300 ml. of ether was added to the reactionmixture to precipitate the salt. The salt was separated by filtration,washed twice with 150 ml. portions of ether and air dried. This salt wasused in the following three reactions.

EXAMPLE 18 The reaction between sodium dithiocarbanilate and allylchloride In a 500 ml., three-necked flask, fitted with a mechanicalstirrer, dropping funnel and thermometer, was placed a. solution of 47.8gm. (0.25 mole) of sodium dithiocarbanilate in 150 ml. of water. To thisconstantly stirred solution, 19.12 gm. (0.25 mole) of allyl chloride wasadded dropwise over a period of twenty minutes. Stirring was continuedand after thirty minutes a white precipitate had separated. Theprecipitate was separated by filtration, washed twice with 150 ml.portions of water and then re-crystallized twice from an acetone-watersolution. The product was then dried in vacuum for eight hours at 50 C.The nitrogen content was found to be 12.27%.

The compound was obviously not an ester of dithiocarbanilic acid. Thenitrogen content and melting point were the same as those ofN,N-diphenyl thiourea and a mixed melting point taken with a knownsample of N,N-diphenyl thiourea was also 154 C. The compound was thuspresumed to be N,N-diphenyl thiourea.

EXAMPLE 19 The reaction between sodium dithiocarbanilate andchloro-Z-propanone In a. 500 ml., three-necked flask, fitted with amechanical stirrer, dropping funnel and thermometer, was placed asolution of 47.8 gm. (0.25 mole) of sodium dithiocarbanilate in 150 ml.of water. To this constantly stirred solution, maintained at 25 C., 23gm. (0.25 mole) of chloro-2-propanone was added dropwise over a periodof 20 minutes. Stirring was continued and after thirty rninutes a tanprecipitate had separated. The precipitate was separated by filtration,washed with three 150 ml. portions of water and then re-crystallizedthree times from an acetone-water solution. The product was then driedin vacuum for forty-eight hours at 40 C. The yield was 40 gm. of acompound melting at 170 C. The nitrogen content was 7.4% as compared to6.2% required for the ester.

The compound was probably not the desired ester of dithiocarbanilic acidbut was kept for antifungal testing.

EXAMPLE 20 The reaction between sodium dithiocarbanilate and ethylbromoacetate In a 500 ml., three-necked flask, fitted with a mechanicalstirrer, dropping funnel and thermometer, was placed a solution of 47.8gm. (0.25 mole) of sodium dithiocarbanilate in 150 ml. of water. To thisconstantly stirred solution, maintained at 25 C., 41.8 gm. (0.25 mole)of ethyl bromoacetate was added dropwise over a period of 30 minutes.Stirring was continued and after one hour a pink metastable oil hadseparated. The mother liquor was poured off the oil and the oil washedthree times with ml. portions of water. The product was then dried invacuum at 40 C. for forty-eight hours during which time crystallizationtook place. The compound was then recrystallized three times from waterand acetone. The yield was 38 gm. of a compound melting at -122 C. Thenitrogen content was found to be 6.4% as compared to 5.5% required forthe ester. The compound was apparently not the desired ester ofdithiocarbanilic acid but was kept for antifungal testing.

The above examples of processes for preparing the condensation reactionproduct of this invention are not intended to limit one to the exactconditions illustrated therein, as it may be readily understood thatvariations in the processes and procedures are possible withouteffecting the results. While in each of the above examples thedimetallic bis-dithiocarbamate is placed in water and has the organichalide added thereto, the water may be replaced in part or completely byalcohol, acetone, benzol, chloroform, etc. Also, while the disodium saltof ethylene bis-dithiocarbamate is used in each of the above examples,other dimetallic bis-dithiocarbamate salts such as potassium, ammonium,lithium, calcium and the like may be used. With sparingly solubledimetallic salts, examples of which are zinc, copper, cadmium and iron,the reaction is much slower and the yield much smaller.

The term organic halide as used herein includes, in addition to thespecific halides disclosed in the above examples, the following: benzoylchloride, acetyl chloride, phenacyl bromide, p-nitro phenacyl bromideand other substituted phenacyl halides, halogenated ketones such aschloro-2-propanone, 3,4-dich1orbenzyl chloride, 2,4-dichlorbenzylchloride, orthochlor benzyl chloride, monochloro acetic acid, monochloropropionic acids, 2,4- dinitro chlorobenzene, picryl chloride, aromaticsulfonyl halides such as benzene sulfonyl chloride, and other radicalscontaining reactive halogens.

The following table illustrates the effect of each of the condensationreaction products formed in Examples 1 to 5 inclusive on particularmicro-organisms and the effect is compared with undecylenic acid, acompound presently in common use as a fungicide for thesemicroorganisms.

TABLE L-WIDTH OF ZONE F INHIBITION IN MILLIMETERS Undee- Bis-2- Bise-Dl-n- Organism ylenie Diallyl Dibenzyl hydroxyhydroxypropyl Acetone acidester ester ethyl propyl ester ester ester 2 42 2 2 2 22 2 27 37 15 1513 so 0 2s 31 15 17 13 2s 0 Aver 25 4 35. 3 16. 19. 5 16. 7 27. 5 0 5260 32 44 31 40 o A. achocnlcinli 46 49 31 33 30 35 0 47 55 25 45 30 35 0Aver 48. 3 54. 7 26.3 40. 7 so. a 35. 7 0 34 53 25 43 41 34 o 35 50 2s47 as as o Aver 35 55. 5 26. 5 45 3s. 5 36 o 50 55 21 46 a2 44 o M.qypseum 45 64 21 55 50 52 0 42 e0 30 55 50 4s 0 Aver 45. 7 53 24 52. 347. 3 4s 0 49 53 2s 49 so 40 o T. rubrum 54 47 25 52 50 40 o 54 55 17 5050 41 0 Aver 52. 3 55 23. 7 50. a 45. 7 4o. 3 o

The esters formed in the remaining examples above 30 (8)Bis(acetylmethyl)-2-hydroxytrimethylene-1, 3-bis-diwere tested on othermicro-organisms for determination thiocarbamate of their fungicidalaction and the results compared with (9) Bis(carbethoxymethyl)-2-hydroxytrimethylene-1, bisundecylenic acid and with acetone. Theseresults are dithiocarbamate set forth in Tables II, III and IV. Thecompounds set Dibenzyl-2-hydroxytrimethylene-1, 3-bis-dithiocarforth andcompared in Tables 11, III and IV are referred 35 bamate to by numberswhich correspond to the following num- (11) Bis (polyethyleneglycol)ethylene bis-dithiocarbambered compounds: ate (aver. M.W. 960) I (1)Bis(2-methylallyl) ethylene bis-dithiocarbamate i ig sg g ggg hylene6646mm561mm-- (2) Bis(acetylmethyl) ethylene bis-dithiocarbamate 13 6 3Bis(3,4-dichlorobenzyl) ethylene bis-dithiocarbamate 4o Reactlon Productof Sodmm dlthwcarbamlate n (4) Bis(p-chlorobenzyl) ethylenebis-dithiocarbamate chlom'z'Propanone (5) Bis(2,4-dichlorobenzyl)ethylene bis-dithiocarbam- Reactlon P c f SOdlllm dithiocarbamlate andate ethyl bromoacetate (6) Bis(forrnylmethyl) ethylenebis-dithiocarbamate Undecylenic acid 45 (16) Acetone (7)Bis(carbethoxymethyl) ethylene bis-dithiocarbamate TABLE II.WIDTH OFZONE OF INHIBITION IN MILLIMETERS, AGENTS AT 5% CONCENTRATION CompoundNumber Organism 35 43 32 24 23 30 32 32 28 34 26 45 40 33 50 0 33 41 2725 24 23 32 32 31 25 25 43 40 35 50 0 M,ca7l,i8 42 35 33 25 24 23 36 4225 25 24 37 35 41 40 o 41 42 31 23 24 25 40 43 32 23 22 38 35 41 45 0 3839 31 23 26 23 38 35 33 32 24 41 40 37 47 0 Aver 37.8 40 30.8 24 24.224.8 35.6 37.8 29.8 27.8 24.2 40.8 38 37.4 46.4 0

- 30 27 30 13 24 18 29 25 32 15 23 26 17 0 28 3O 13 24 18 27 25 33 20 1825 26 18 25 0 M,gypseum 27 31 25 16 19 20 34 24 21 13 27 27 24 32 27 032 28 20 16 18 20 37 23 20 13 21 25 25 32 0 3O 30 22 15 18 19 44 24 2318 19 35 25 21 0 Aver 29.4 29.2 25.4 14.6 20.6 19 34.2 24.2 25.8 16.8 2027 25 24 30.4 0

21 28 14 13 15 15 16 16 14 13 13 13 13 16 28 0 27 32 14 13 15 15 16 1714 13 13 13 13 16 30 0 M,audouim' 23 34 13 13 14 15 17 17 15 13 13 13 1315 27 0 [25 35 15 13 16 15 15 16 13 13 13 13 13 17 29 0 25 31 13 13 1515 17 16 15 13 13 13 13 16 32 0 Aver 24.2 32 13.8 13 15 15 16.2 16.414.2 13 13 13 13 16 29.2 0

38 25 13 23 23 48 29 33 39 39 38 30 38 60 0 35 58 25 13 28 23 48 28 3135 80 47 30 38 60 O T. mentagrophytes 29 60 36 15 30 28 28 39 35 35 4046 20 30 60 0 29 36 16 35 25 40 38 35 39 40 40 20 30 60 0 40 60 35 18 2527 42 29 33 40 35 40 26 36 60 0 Aver 34.2 60.6 31.4 15 27.2 25.2 41.232.6 33.4 37.6 36.8 42.2 25.2 34.4 60 0 AT 2%% CONCENTRATION CompoundNumber Organism 32 32 20 25 35 0 3O 21 35 25 19 29 0 M,cania 28 17 24 2425 35 0 30 16 35 22 19 22 28 0 28 18 34 23 21 24 29 0 Average 29. 6 18.4 34. 2 23. 8 20. 6 25. 2 31. 2 O

28 20 30 20 20 20 32 0 2% 23 i3 i2 2 32 2 8 umn M WP 23 19 37 17 19 2531 0 30 21 33 20 20 25 33 0 Average 26. 4 21 34. 8 19 20. 2 24. 2 29. 80

13 13 13 13 13 13 16 0 14 15 13 13 13 15 17 0 M. audouini 3 17 13 13 1314 14 0 14 16 13 13 13 14 14 0 14 15 13 13 13 13 16 0 Average 13.6 15.213 13 13 13.8 15.4 0

35 28 20 13 20 25 36 0 h u 33 3% 35 ii ii 2 32 3 mm! m 1/ 8 T M p 35 3125 16 19 23 37 0 30 21 14 21 24 40 0 Average 38. 6 29. 4 22. 4 14. 8 2024. 2 35. 6 0

% CONCENTRATION Compound Number Organism 27 15 a2 17 19 15 1s 0 27 1a 351a 19 17 17 o me 30 17 33 13 1s 13 13 o 30 16 a0 16 2o 13 1a 0 2s 17 a414 21 1s 13 0 Average 28.4 15.6 32.8 14.6 19.4 15.2 14.3 o

36 1s 23 13 1a 1a 20 0 33 58 it it i? ii it a um 0 M c 23 17 .20 15 1716 21 0 26 1s 21 17 16 1a 24 0 Average 25.2 1&4 21.8 15.2 14.8 14.4 21.20

1a 1a 1a 1a 1a 13 1a 0 13 1a 1a 13 1a 1a 13 o M,u,dou,ini 1a 13 13 13 131a 13 o 13 1a 13 1a 1a 1a 1a 0 13 1a 13 1a 13 1a 1a 0 Average 13 13 1313 13 13 13 0 '35 29 19 1a 1a 1a 25 o 37 26 22 1a 14 13 2s 0 T.nuntavrophvtea 34 30 25 13 15 13 30 0 33 27 21 13 1a 1a 23 0 a4 29 21 1a14 13 2a 0 Average 34.6 28.2 21.6 13 13.8 13 25.8 6

The C t medium Used in Obtaining the results in one milliliter of thewater suspensions of the spores was the above tables was Sabouraudsdextrose agar main- 60 streaked on each plate. All plates in a serieswere inoctained at a pH of 5.6 to prevent bacterial contamination.ulated from the same spore suspension so as to insure Wa e Suspensions0f spores of the test Organlsms a nearly uniform number of spores ineach plate. The indicated in the tables were prepared from 14 to 21 daypetri dishes, when ready for use, thus contain a thin upper Sabouraudsdextrose agar slant cultures which had been layer in which the funguscells were embedded and a 20 incubated at 28 C. The suspensions wereprepared by milliliter base layer below. grinding the growing coloniesin the slant tube with a To test each particular fungicide sterilefilter paper ground-glass rod. Fifteen milliliters of sterile water fordisks, 13 mm. in diameter, were placed in the acetone injection at 60 C.were then added and suspension of solution containing the fungicide.These disks were then the spores brought about by vigorous agitation.The picked up with sterile forceps, and the excess liquid antifungalagents were prepared for use in the experiment drained off by touchingthe disk to the wall of the conby making up weight to volume solutionsof each agent in acetone.

The method of testing consisted of pipetting 20 ml. of the culturemedium into a sterile petri dish and allowing tainer. The disk was thenimmediately dropped onto the agar surface. After a constant incubationat 37 C., for seven days the diameter of the zones of inhibition weremeasured and recorded. A control plate for each series the medium toharden. After solidification of the agar, using acetone was included inall of the observations.

13 The results of these experiments are recorded in Tables II, III andIV.

The bis-dithiocarbamate esters formed in the above examples range from alight yellow nearly odorless solid with the dibenzyl ester to a ratherthick yellow oil-like compound of garlic odor with the diallyl ester.The range of properties of these compounds permits their use in a widevariety of fungicidal, germicidal, bactericidal and antiseptic types ofpreparations and also as preservative agents. They may be used in solidform, in solution, in emulsion and combined with other active or inertingredients to make sprays and preservative paints. The condensationproducts, namely, the esters of the bisdithiocarbamates may bepracticably applied to the desired surfaces in solutions or suspensionsmade in alcohol, water, alcohol and water, acetone, acetone and water,isopropyl alcohol, isopropyl alcohol and water, propylene glycol, andother forms. Such solutions or suspensions may be used as fungicidalpreparations to disinfect inanimate objects.

These new esters of bis-dithiocarbamate also have the advantage of beingmore stable than the corresponding dimetallic salts. The esters showonly slight or no decomposition over periods ranging up to two yearswhile the dimetallic salts may show considerable decomposition in threeto six months. The dimetallic salts have been reported to be quiteunstable in aqueous solutions and lose up to ninety percent of theiractivity in a short time. Aqueous suspension of the esters of thisinvention have given no evidence of decomposition. These condensationproducts are also advantageous in that they are fungicidal, as shown bythe fact that transplants of media taken from zones of inhibition causedby the esters and fungus growth will not grow, whereas fatty acids, suchas undecylenic acid are only weakly fungistatic.

It is to be understood that various changes, modifications, anddepartures from this specific disclosure may be made by those skilled inthe art without departing from the scope and spirit of the invention asdescribed herein and as defined in the appended claims.

Having fully described the invention, What is claimed 1. The process ofinhibiting and destroying microorganisms selected from the groupconsisting of Microsporum audom'ni, M. gypseum, M. lanosum, M. canis, Trichophyton schoenleini, T. rubrum, T. mentagrophytes, andStaphylococcus aureus comprising applying to the micro-organisms afungicidal amount of alkylene bisdithiocarbamate ester.

2. The process as defined in claim 1 wherein said ester is di-allylethylene bis-dithiocarbamate.

3. The process as defined in claim 1 wherein said ester isbis-Z-hydroxyethyl ethylene bis-dithiocarbarnate.

4. The process as defined in claim 1 wherein said ester is bis-(2 methylallyl)-ethylene bis-dithiocarbamate.

5. The process as defined in claim 1 wherein said ester isbis-acetylmethyl ethylene bis-dithiocarbamate.

6. The process as defined in claim 1 wherein said ester isbis-(carbethoxymethyl) ethylene bis-dithiocarbamate.

References Cited in the file of this patent UNITED STATES PATENTS Re.23,742 Hester Nov. 24, 1953 2,313,871 Hanford et a1 Mar. 16, 19432,561,208 Kirk July 17, 1951 2,796,376 Williams et a1 June 18, 1957OTHER REFERENCES Zinsser: Bacteriology, 11th ed., Appleton-CenturyCrofts, Inc., N.Y., 1957, pp. 873-880.

Collins: Synthesis and Invest. of Some Bis-Dithiocarbarnate Esters andPossible Fungicides, deposited in Lib. of Iowa State Univ., February1954 (35 pp.).

1. THE PROCESS OF INHIBITING AND DESTROYING MICROORGANISMS SELECTED FROMTHE GROUP CONSISTING OF MICROSPORUM AUDOUINI, M. GYPSEUM, M. LANSOUM, M.CANIS, TRICHOPHYTON SCHOENLEINI, T. RUBRUM, T. MENTAGROPHYTES, ANDSTAPHYLOCOCCUS AUREUS COMPRISING APPLYING TO THE MICRO-ORGANISMS AFUNGICICDAL AMOUNT OF ALKYLENE BISDITHIOCARBAMATE ESTER.